Amidase (gene TAM) from Thauera sinica K11

Enzyme Description

Species

Extremophile

EC Number

3.5.1.4 ↗

Sequence

Length: 371 amino acids

MSIFVVKLDLGGSGPTVAIKDTIDVAGVPTRAGSRAFEDAAPAAANAAVVDRLLAAGYHLVGKANMHELAFGTTGLNAWTGTPVNPRHPAYVPGGSSSGSAAAVAAGLAEAALGTDTGGSIRIPAACCGVFGLKPTFGRVSRDGVMPAQTSLDCVGPFAGDMDALIACMRAIDPDFGALPDTAGVTVGLVEVEARPEIRRAVEAAVAAGGFAAGRVVLDAFVRAYDAGLVVINAETWAACGELVATGRVGEDVAKRLLAASRTTAADVQAAEAVRAAFTAQVDEALRTFPILALPTMADAPPLLESGADTSKLVGMTSLVRPFNLSGHPAIAIPLQGAGDFPMSLQLVAAKGRDELLCAVARQIAGRLDQH

Wenfei Tan et al. (2021) — Cloning, expression and biochemical characterization of a novel amidase from Thauera sinica K11

Protein Expression and Purification · doi:10.1016/j.pep.2020.105751 ↗ · Stability - Incubation

6 measurements ▼

Database ID

UniProt: A0ABW1AP76 ↗

Sequence Annotation

Explicit - Provided

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (6 measurements)

6 measurements

Property	Assay	Solvent	Solvent Volume	Aqueous Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (Brammar-Clarke method, ferric–butoxyhydroxamate complex absorbance mesasurement, 500 nm) in aqueous phase	Methanol	5%	100.0	32.8	%	Incubation: ?, Assay: 100 mM Tris-HCl buffer	Incubation: ?, Assay: 7.5	Incubation: 4°C, Assay: 45°C	0.1 M Butyramide, 1 M Hydroxylamine	Butoxyhydroxamic acid , NH3	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (Brammar-Clarke method, ferric–butoxyhydroxamate complex absorbance mesasurement, 500 nm) in aqueous phase	Ethanol	5%	100.0	36.3	%	Incubation: ?, Assay: 100 mM Tris-HCl buffer	Incubation: ?, Assay: 7.5	Incubation: 4°C, Assay: 45°C	0.1 M Butyramide, 1 M Hydroxylamine	Butoxyhydroxamic acid , NH3	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (Brammar-Clarke method, ferric–butoxyhydroxamate complex absorbance mesasurement, 500 nm) in aqueous phase	Acetonitrile	5%	100.0	32.2	%	Incubation: ?, Assay: 100 mM Tris-HCl buffer	Incubation: ?, Assay: 7.5	Incubation: 4°C, Assay: 45°C	0.1 M Butyramide, 1 M Hydroxylamine	Butoxyhydroxamic acid , NH3	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (Brammar-Clarke method, ferric–butoxyhydroxamate complex absorbance mesasurement, 500 nm) in aqueous phase	Acetone	5%	100.0	5.1	%	Incubation: ?, Assay: 100 mM Tris-HCl buffer	Incubation: ?, Assay: 7.5	Incubation: 4°C, Assay: 45°C	0.1 M Butyramide, 1 M Hydroxylamine	Butoxyhydroxamic acid , NH3	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (Brammar-Clarke method, ferric–butoxyhydroxamate complex absorbance mesasurement, 500 nm) in aqueous phase	Isoamyl Alcohol	5%	100.0	12.8	%	Incubation: ?, Assay: 100 mM Tris-HCl buffer	Incubation: ?, Assay: 7.5	Incubation: 4°C, Assay: 45°C	0.1 M Butyramide, 1 M Hydroxylamine	Butoxyhydroxamic acid , NH3	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (Brammar-Clarke method, ferric–butoxyhydroxamate complex absorbance mesasurement, 500 nm)	Dimethyl Sulfoxide (DMSO)	5%	100.0	85.8	%	Incubation: ?, Assay: 100 mM Tris-HCl buffer	Incubation: ?, Assay: 7.5	Incubation: 4°C, Assay: 45°C	0.1 M Butyramide, 1 M Hydroxylamine	Butoxyhydroxamic acid , NH3	—	—	Non-incubated control (in %), assay in aqueous phase

Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (Brammar-Clarke method, ferric–butoxyhydroxamate complex absorbance mesasurement, 500 nm) in aqueous phase	Methanol	5%	100.0	32.8	%	Incubation: ?, Assay: 100 mM Tris-HCl buffer	Incubation: ?, Assay: 7.5	Incubation: 4°C, Assay: 45°C	0.1 M Butyramide, 1 M Hydroxylamine	Butoxyhydroxamic acid, NH3	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (Brammar-Clarke method, ferric–butoxyhydroxamate complex absorbance mesasurement, 500 nm) in aqueous phase	Ethanol	5%	100.0	36.3	%	Incubation: ?, Assay: 100 mM Tris-HCl buffer	Incubation: ?, Assay: 7.5	Incubation: 4°C, Assay: 45°C	0.1 M Butyramide, 1 M Hydroxylamine	Butoxyhydroxamic acid, NH3	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (Brammar-Clarke method, ferric–butoxyhydroxamate complex absorbance mesasurement, 500 nm) in aqueous phase	Acetonitrile	5%	100.0	32.2	%	Incubation: ?, Assay: 100 mM Tris-HCl buffer	Incubation: ?, Assay: 7.5	Incubation: 4°C, Assay: 45°C	0.1 M Butyramide, 1 M Hydroxylamine	Butoxyhydroxamic acid, NH3	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (Brammar-Clarke method, ferric–butoxyhydroxamate complex absorbance mesasurement, 500 nm) in aqueous phase	Acetone	5%	100.0	5.1	%	Incubation: ?, Assay: 100 mM Tris-HCl buffer	Incubation: ?, Assay: 7.5	Incubation: 4°C, Assay: 45°C	0.1 M Butyramide, 1 M Hydroxylamine	Butoxyhydroxamic acid, NH3	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (Brammar-Clarke method, ferric–butoxyhydroxamate complex absorbance mesasurement, 500 nm) in aqueous phase	Isoamyl Alcohol	5%	100.0	12.8	%	Incubation: ?, Assay: 100 mM Tris-HCl buffer	Incubation: ?, Assay: 7.5	Incubation: 4°C, Assay: 45°C	0.1 M Butyramide, 1 M Hydroxylamine	Butoxyhydroxamic acid, NH3	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (Brammar-Clarke method, ferric–butoxyhydroxamate complex absorbance mesasurement, 500 nm)	Dimethyl Sulfoxide (DMSO)	5%	100.0	85.8	%	Incubation: ?, Assay: 100 mM Tris-HCl buffer	Incubation: ?, Assay: 7.5	Incubation: 4°C, Assay: 45°C	0.1 M Butyramide, 1 M Hydroxylamine	Butoxyhydroxamic acid, NH3	—	—	Non-incubated control (in %), assay in aqueous phase

Visualization : Stability — Incubation

One bar per measurement. Colour = solvent, shade = solvent volume.

Structure

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